System and methods for remote assessment of a sample assay for disease diagnostics

ABSTRACT

A method for performing a lateral flow assay is provided. The method includes inserting a sample cartridge ( 201 ) in a dark chamber ( 220 ) and activating a light emitter ( 251 ) in the dark chamber ( 220 ). The method includes focusing an optical coupling mechanism ( 115   a,    115   b ) in an image-capturing device ( 100   a,    100   b ) to optimize an image of a sensitive area ( 202 ) in the sample cartridge ( 201 ) and capturing, with an image capturing device ( 100   a,    100   b ), an image of a sensitive area ( 202 ) in the sample cartridge ( 201 ) after a selected period of time. The method also includes providing the image of the sensitive area ( 202 ) to a processor, wherein the processor comprises an image-capturing application ( 122 ). A system and a computer-implemented method to perform at least partially the above method are also provided.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional Application No.62/792,813, filed on Jan. 15, 2019 and of U.S. Provisional ApplicationNo. 62/843,846, filed on May 6, 2019, each incorporated by referenceherein in its entirety.

TECHNICAL FIELD

The present disclosure generally relates to devices and methods fortelemedicine applications and other in-situ immunoassay measurements.More specifically, the present disclosure relates to consumables, whichin conjunction with client devices used by consumers may be used in asimple and accurate procedure to assess a disease diagnostic locallyand/or remotely.

BACKGROUND

Currently, disease diagnostics using test assays involve users sendingtest samples to a laboratory for accurate analysis. This step istime-consuming, as it involves the physical displacement of a samplecartridge back (with the test sample) and forth (before use of the testsample) between the medical provider (e.g., clinic, physician, pharmacy,or laboratory), and the user. Furthermore, these test samples tend todrag the queues in the list of tasks of clinical laboratories, manytimes unnecessarily (as some samples, or most, may be negative).Additionally the need to visit a doctor's office in order to have thetest performed may help the spread of infectious diseases through theexposure of uninfected patients to the carrier of a positive test (e.g.,in the waiting room of a doctor's office). Further, the time lag betweentest and result may be a potential hazard, e.g., for epidemic orpandemic emergencies, or when the outcome of treatment of a seriouscondition is dramatically impacted by the time of start of a therapy.

BRIEF SUMMARY

In a first aspect, a method comprising inserting a sample cartridge in adark chamber and activating a light emitter in the dark chamber isprovided. The method also comprises focusing an optical couplingmechanism in an image-capturing device to optimize an image of asensitive area in the sample cartridge, capturing, with an imagecapturing device, an image of a sensitive area in the sample cartridgeafter a selected period of time, and providing the image of thesensitive area to a processor, wherein the processor comprises animage-capturing application.

In another aspect, a system is provided that comprises an enclosureincluding a dark chamber, the enclosure configured to block ambientlight from entering the dark chamber. The system also comprises acartridge aperture on a side of the enclosure to enable a samplecartridge to be disposed at least partially inside the dark chamber,wherein the cartridge aperture is configured to at least partially closewhen the sample cartridge is at least partially disposed inside the darkchamber. The system also comprises an optical coupler inside theenclosure to form a partial image of the sample cartridge in a sensorarray of an image-capturing device, when the sample cartridge is atleast partially disposed in the dark chamber. The system also comprisesa light emitting device in an interior portion of the enclosure, thelight emitting device configured to emit a fluorescence excitation lightdirected to the sample cartridge.

In another aspect, a computer-implemented method is provided thatcomprises identifying, upon receipt of a user input, a fiduciary figurein a bottom side of an enclosure with an image-capturing device andadjusting, in the image-capturing device, an optical coupling to obtaina sharp image of the fiduciary figure. The computer-implemented methodalso comprises identifying a sensitive area of a sample cartridge withina field of view of the optical coupling and finding a border of thesensitive area of the sample cartridge and applying geometricaltransformations on an area delimited by the border of the samplecartridge. The computer-implemented method also comprises identifying atarget region within the sensitive area of the sample cartridge,extracting a value of a selected color for multiple pixels in the targetregion, and determining a presence of a target analyte when the value ofthe selected color is greater than a pre-selected threshold.

BRIEF DESCRIPTION OF THE FIGURES

FIGS. 1A-1B illustrate architectures including a remote server, adatabase, and an image-capturing device to collect an image from asample cartridge in an enclosure, according to some embodiments.

FIG. 2 illustrates an enclosure including a cartridge aperture toreceive a sample cartridge, according to some embodiments.

FIGS. 3A-3D are block diagrams illustrating steps in a method for remoteassessment of a sample assay for disease diagnostics, according to someembodiments.

FIG. 4A illustrates an enclosure including a sample cartridge having acolored fiducial for identifying the location of a sample assay feature,according to some embodiments.

FIG. 4B illustrates an enclosure including a sample cartridge having afluorescent fiducial for identifying the location of a sample assayfeature, according to some embodiments.

FIG. 5 illustrates a series of images of the sensitive area of a samplecartridge including a blank assay, collected over time, according tosome embodiments.

FIG. 6 illustrates an image of a sensitive area of a sample cartridgeincluding a sample assay and positioning marks measured with animage-capturing device, according to some embodiments.

FIG. 7 illustrates an image of a sensitive area of a sample cartridgecollected and processed by an image-capturing device, according to someembodiments.

FIG. 8 illustrates cross-sections of fluorescence signals captured froma sample assay along a longitudinal direction, according to someembodiments.

FIG. 9 is a flow chart illustrating steps in a method for capturing animage of a sensitive area in a sample cartridge using an image-capturingdevice and a dark chamber, according to some embodiments.

FIG. 10 is a flow chart illustrating steps in a computer-implementedmethod for remotely diagnosing a disease with an image-capturing device,according to some embodiments.

FIG. 11 is a flow chart illustrating steps in a method for remotelydiagnosing a disease with an image-capturing device, according to someembodiments.

FIG. 12 is a flow chart illustrating steps in a method for diagnosing adisease with an image-capturing device, according to some embodiments.

FIG. 13 is a flow chart illustrating steps in a method for diagnosing adisease with an image-capturing device, according to some embodiments.

FIG. 14 is a block diagram illustrating an example computer system withwhich the image-capturing device and the server of FIGS. 1A-1B, andmethods as disclosed herein can be implemented, according to someembodiments.

DETAILED DESCRIPTION

In the following detailed description, numerous specific details are setforth to provide a full understanding of the present disclosure. It willbe apparent, however, to one ordinarily skilled in the art, that theembodiments of the present disclosure may be practiced without some ofthese specific details. In other instances, well-known structures andtechniques have not been shown in detail so as not to obscure thedisclosure.

In the burgeoning area of telemedicine, it has become increasinglydesirable to take advantage of the almost universal availability ofelectronic appliances that may have wireless network access and sensorsand that may also include increasingly higher computationalcapabilities. The image-capturing capabilities of mobile computerdevices and other consumer products have increased resolution and offerthe capability for digital processing (e.g., spatial filtering andadjustment, and spectral filtering). There remains a growing need forremote measurement of immunoassays for the detection of chemical andbiological agents or pathogens may include security tests and screening(e.g., at airports, police, and military checkpoints), or environmentalanalysis and monitoring (e.g., air pollution, contamination of waterways and reservoirs—for disease control or agricultural production-, andthe like).

Embodiments consistent with the present disclosure take advantage of thehigh image-capturing and processing capabilities of consumer appliancesto provide simple yet accurate diagnostic method, system and procedurefor detection of, for example, infectious agents or biomarkers ofdiseases or disorders (e.g., legionella, influenza, Ebola, Lyme disease,and the like). The types of tests consistent with embodiments in thepresent disclosure may include any type of spectroscopic analysis oftest assays using electromagnetic radiation, such as, withoutlimitation, absorption spectroscopy (ultra-violet, visible, or infrared)including reflectance or transmittance spectroscopy, or emissionspectroscopy, including fluorescence and luminescence spectroscopy,Raman spectroscopy, and any type of radiation scattering. Moreover,embodiments as disclosed herein may further exploit the networkingcapabilities of such appliances to enhance the processing capabilitiesof each test by using cloud-computing solutions. Accordingly, in someembodiments, a high quality (e.g., high spatial and spectral resolution)image, sequence of images, or video is uploaded to a remote server thatcan perform massively parallel computations to provide, in a reducedtime, a diagnostic result. Such analyzed material may be processedimmediately, at a later date/time, and or may be compared to previouslycollected materials to determine differences over time, e.g., a timeevolution of the analyte across a test strip.

The subject system provides several advantages, including the abilityfor a user to quickly learn whether a disease is present or latent,without the need to access specialized personnel, or a complex machineor instrument.

Some embodiments provide the advantage of widely broadening the marketfor medical test kits, as consumers who have wide access toimage-capturing devices in the form of mobile computing devices andother appliances, may desire to perform tests even before perceiving anysymptoms or going to a doctor or clinic. This also may provide theadvantage of a screening step before people attend clinics to avoidexposure of innocent bystanders, or saturate the resources of a givenmedical facility. Further, the cost of a test for a remote user ofmethods as disclosed herein may be substantially lower than the costassociated with a visit to a clinic or laboratory, including waitingtimes and scheduling.

The proposed solution further provides improvements to the functioningof computers (e.g., the server or a user mobile device) because it savesdata storage space and interaction time by enabling a remotetransmission of image analysis data and results (e.g., pictures,sequences of pictures, and/or videos).

Although many examples provided herein describe a user's personalinformation and data as being identifiable, or a download and storage ofa user interaction history with one or more remote clinics, each usermay grant explicit permission for such user information to be shared orstored. The explicit permission may be granted using privacy controlsintegrated into the disclosed system. Each user may be provided noticethat such user information will be shared with explicit consent, andeach user may at any time end the information sharing, and may deleteany stored user information. Further, in some embodiments, the storedand/or transmitted user information may be encrypted to protect usersecurity and identity.

FIG. 1A illustrates an architecture 10A including a remote server 130, adatabase 152, and an image-capturing device 100A to collect an image orvideo from a sample cartridge 101 in an enclosure 120A, according tosome embodiments. In some embodiments, database 152, server 130, andimage-capturing device 100A may be communicatively coupled via a network150 (e.g., through an Ethernet link, an optical link, a wireless link, acellular network, and the like). Network 150 can include, for example,any one or more of a local area network (LAN), a wide area network(WAN), the Internet, and the like. Further, network 150 can include, butis not limited to, any one or more of the following network topologies,including a bus network, a star network, a ring network, a mesh network,a star-bus network, tree or hierarchical network, and the like.

In the architecture illustrated in the figure, sample cartridge 101 andenclosure 120A may be consumables that the user may dispose of afteruse. For example, sample cartridge 101 may be discarded or disposed ofafter a single use with a test sample, while enclosure 120A may be usedmore than one time, and in some embodiments is used repeatedly. In thatregard, sample cartridge 101 and enclosure 120A may be part of a packageor kit requested by or provided to the user from a clinical serviceprovider. The package may include one enclosure 120A and multiplecartridges 101 that may be used with it. Alternatively, enclosure 120Amay be provided separately from the cartridge(s) 101.

In one embodiment, remote diagnostic architecture 10 a includes animage-capturing device 100 a provided by the user. This may include asmartphone or other mobile computing device (e.g., tablet, pad, or evenlaptop) including a sensor array 140 and an optics coupling mechanism115A (e.g., a lens system with autofocus capabilities). Image-capturingdevice 100 a may be configured to couple wirelessly, through a network,with remote server 130 and database 152. Remote server 130 may providesupport for an image-capturing application 122 installed inimage-capturing device 100 a. The support may include updateinstallation, retrieval of raw data (e.g., pictures, sequences ofpictures and videos) for storage in database 152, image processing, andthe like.

FIG. 1B illustrates an architecture 10 b including a remote server 130,a database 152, a user device 110, and an enclosure 120 b to collect animage or video from sample cartridge 101, according to anotherembodiment. Client device 110 may include a smartphone or other mobilecomputing device (e.g., tablet, pad, or even laptop). Accordingly, insome embodiments, architecture 10B includes a user of client device 110who has ordered a kit including sample cartridge 101 and enclosure 120Band is ready to perform a personal test for a disease or conditionremotely from a hospital or clinic (e.g. at home, in a pharmacy, or inany other location).

In architecture 10B, enclosure 120B includes an image-capturing device100B and wirelessly transmits an image of sample cartridge 101 to clientdevice 110. Client device 110 then may transmit the image or video to aremote server 130, to database 152, or both, via network 150, forprocessing. In some embodiments, client device 110 may perform at leastone or more operations to the image or one or more image frames from avideo before transmitting the image to server 130 or to database 152.For example, in some embodiments, client device may perform at least oneor more quality control steps over the one or more images provided byenclosure 120B before transmitting to server 130. In some embodiments,client device 110 may obtain a preliminary or a definitive diagnosticbased on the analysis of the image of sample cartridge 101. Accordingly,in some embodiments, client device 110 may transmit the preliminary ordefinitive diagnostic to server 130 with or without an image of samplecartridge 101.

Client device 110 communicates with server 130 and with enclosure 120Busing a communications module 118-1. Enclosure 120B may communicate withclient device 110 through a communications module 118-2. Communicationsmodules 118-1 and 118-2 will be collectively referred to, hereinafter,as “communications modules 118.” Communications modules 118 may includehardware and software associated with radio-frequency (RF) antennas forcommunication via WiFi, Bluetooth (e.g., low energy Bluetooth, BLE), ornearfield contact (NFC) protocols. For example, when enclosure 120B andclient device 110 are relatively close to each other, communicationsmodule 118 may include a BLE or NFC protocol.

In some embodiments, image-capturing device 100 b in enclosure 120 b mayinclude a sensor array 140 and an optics coupling mechanism 115 b (e.g.,a lens system with autofocus capabilities). Sensor array 140 may collectone or more images of sample cartridge 101 at a desired frame rate, toform a video. In some embodiments, sensor array 140 may collect a singleimage of sample cartridge 101 (e.g., after an assay has run its course),or more than one image (e.g., before and after an assay runs itscourse). In yet some embodiments, sensor array 140 may collect multipleimages of sample cartridge 101 at a pre-selected frequency rate. Thefrequency rate may be adjusted, modified, accelerated, or slowed, basedon preliminary or quality control tests performed by client device 110.In some embodiments, sensor array 140 is coupled to a processor that mayperform, at least partially, an analysis of an image before transmittingit to client device 110.

Remote server 130 may provide support for an image-capturing application122 installed in client device 110. The support may include updateinstallation, retrieval of raw data (e.g., pictures, sequences ofpictures and videos) for storage in database 152, image processing, andthe like. Image-capturing application 122 may include commands andinstructions to control image-capturing device 100 b. Image-capturingapplication 122 may also include commands and instructions to perform atleast a partial analysis of the one or more images provided by enclosure120B. For example, in some embodiments, the instructions inimage-capturing application 122 may include a neural network (NN),artificial intelligence (AI), or machine learning (ML) algorithm toassess a diagnostic based on the one or more images of sample cartridge101. Additionally, in some embodiments, image-capturing application 122may include instructions to assess a quality control of the one or moreimages provided by enclosure 120B, based on sensor data indicative ofthe positioning of sample cartridge 101 within enclosure 120B. Thesensor data may be provided by sensors disposed within enclosure 120B.

In some embodiments, device 110 may further include an image-capturingdevice 100 c to collect an image of a fiduciary label 105 on samplecartridge 101. Accordingly, image-capturing application 122 mayincorporate the image of fiduciary label 105 on sample cartridge 101into a measurement protocol. The measurement protocol may be transmittedby device 110 to server 130 and/or to database 152, where metadataassociated with sampling cartridge 101 may be correlated withinformation stored therein. For example, in some embodiments, themetadata in fiduciary label 105 may be correlated with a user ID and/orwith an assay identification code (e.g., an indicator of the assay onsample cartridge 101, such as an immunoassay for influenza A and/or B,Streptococcus pneumoniae, Group A Streptococcal, Group B Streptococcal,respiratory syncytial virus (RSV), herpes simplex virus (HSV 1 and/orHSV 2), human metapneumovirus (hMPV), Legionella pneumophilia, or otherinfectious agent; an assay for antibodies associated with Lyme disease;an assay for markers of pregnancy (human chorionic gonadotropin),sexually transmitted disease (e.g., Chlamydia), hepatitis, or any otherdisease or condition).

In some embodiments, image-capturing application 122 may also includeinstructions for the user as to the mode of use and a measurementprotocol for sample cartridge 101. For example, the instructions mayillustrate to the user, step by step, how to collect a sample (e.g.,using a swab or other extraction mechanism), mix the sample withappropriate reagents, and provide at least a portion of the sample intosample cartridge 101. Accordingly, image-capturing application 122 maydisplay the instructions and other illustrative icons to the user on adisplay 116 of client device 110.

Hereinafter, architectures 10 a and 10 b will be collectively referredto as “architectures 10,” image-capturing devices 100 a and 100 b willbe collectively referred to as “image-capturing devices 100,” enclosures120 a and 120 b will be collectively referred to as “enclosures 120 oras “dark chambers” 120,” and optics coupling mechanisms 115 a and 115 bwill be collectively referred to as “optics coupling mechanisms 115.”

FIG. 2 illustrates an enclosure 220 (also referred to as a dark chamber)including a cartridge aperture 212 to receive a sample cartridge 201,according to some embodiments. In some embodiments, enclosure 220 has aflat top side and a flat bottom side. In some embodiments, cartridgeaperture 212 is disposed on a side adjacent to the flat bottom side andis configured to enable sample cartridge 201 to be disposed at leastpartially on the flat bottom side that is facing the flat top side.Cartridge aperture 212 is configured to at least partially close whensample cartridge 201 is at least partially disposed on the flat bottomside, on an interior portion of enclosure 220. In this manner, any straylight exterior to the enclosure is prevented from reaching a sensorarray in the image-capturing device (e.g., sensor array 140 inimage-capturing devices 100, cf. FIGS. 1A-1B).

An image capture aperture 225, preferably disposed on a side of theenclosure such as the top side, enables an optical coupling mechanism inthe image-capturing device, when the image-capturing device is disposedon an exterior portion of the enclosure or within the interior of theenclosure (e.g., optics coupling mechanisms 115 in architectures 10, cf.FIGS. 1A-1B). Thus, the image-capturing device may form a partial imageof sample cartridge 201 when sample cartridge 201 is at least partiallydisposed on the flat bottom side. A light emitting device 251 in aninterior portion of enclosure 220 may emit a fluorescence excitationlight 255. Accordingly, light emitting device 251 is configured toexcite a fluorescence light 260 from sample cartridge 201. Further, insome embodiments, light emitting device 251 is selected so thatfluorescence light 260 has a wavelength within a selected color in thesensor array of the image-capturing device. For example, in someembodiments, light emitting device 251 is configured to emit a light ata wavelength of about 385 nm (ultra-violet) to excite a fluorescentradiation from a fluorophore at about 650 nm. Other combinations ofexcitation wavelengths and fluorescence radiation wavelengths may beused according to the convenience of finding adequate fluorophoresubstances and compounds. In some embodiments, the wavelengthcombination of fluorescence excitation light 255 and the fluorescenceemission light 260 is based, at least partially, on the sensitivity andspectral resolution of the sensor array in the image-capturing device.

In some embodiments, enclosure 220 also includes a battery 261configured to provide a power to light emitting device 251. Enclosure220 further includes a circuit 262 coupling battery 261 with lightemitting device 251. In some embodiments, circuit 262 is configured toremain open (e.g., light emitting device 251 ‘off’) when samplecartridge 201 is outside enclosure 220, and to be closed (e.g., lightemitting device 251 ‘on’) when sample cartridge 201 is disposed at leastpartially on the flat bottom side. In some embodiments, circuit 262includes a switch 263 to enable and/or disable light emitting device251. In some embodiments, insertion of battery 261 can cause lightemitting device 251 to function. Battery 261 may include a rechargeablebattery, or a disposable battery. Further, in some embodiments, a powersupply for light emitting device 251 may be provided by theimage-capturing device, via a wired or a wireless connection (e.g., viacommunications modules 118, cf. FIG. 1B).

In some embodiments, a focusing mark 207 is positioned in the interiorof the enclosure. For example, FIG. 2 shows a focusing mark 207 on theflat bottom side of enclosure 220 on the interior portion. Focusing mark207 is configured to allow the image-capturing device to focus theoptical coupling mechanism at the appropriate focal distance 222 whenlight emitting device 251 is turned ‘on.’ In some embodiments, focusingmark 207 may include a two-dimensional pattern to serve as a guide inthe image-capturing application (e.g., image-capturing application 122,cf. FIGS. 1A-1B) of the image-capturing device to rotate and/or de-skewthe image and find the correct orientation of enclosure 220 relative tothe image-capturing device. In some embodiments, sample cartridge 201may include a fiduciary tag 205 that the image-capturing device may alsouse as a focusing mark to further identify the relative orientation andpositioning of a sensitive area 202 relative to the image-capturingdevice. Fiduciary tag 205 may also include information readable by theimage-capturing device, indicative of a type of test assay included inthe sensitive area, dimensions and other characteristics of thesensitive area that may be relevant for the test assessment, includingan identification of the user and or patient providing the sample fortest. Thus, fluorescent emission light 260 may be collected throughimage-capturing aperture 225 by the optics coupling mechanism of theimage-capturing device.

While some of the descriptions herein are focused on fluorescencespectroscopic analysis of the sample cartridge, some embodimentsconsistent with the present disclosure may include any other type ofelectromagnetic interaction and spectroscopic analysis. Some examples ofspectroscopic analysis consistent with the present disclosure mayinclude Raman spectroscopy, infrared absorption spectroscopy, infraredreflectance/transmittance spectroscopy, and the like. Furthermore, insome embodiments, light emitting source 251 may be replaced by anoptical coupling mechanism (e.g., a lens, mirror, prism, diffractiongrating, or any combination thereof) to use solar radiation (e.g.,during day light) or any exterior illumination to excite a spectroscopicresponse of the sensitive area in sample cartridge 201.

Enclosure 220 is configured to avoid or control any external light tointerfere with fluorescence excitation light 255 or with thefluorescence emission light 260 collected by the image-capturing device.For example, it is desirable to illuminate sensitive area 202 in samplecartridge 201 uniformly (e.g., no shadows, bright spots, or otherartifacts) to create a smooth spectroscopic background that can befiltered out by the image-capturing application in the image-capturingdevice.

FIGS. 3A-3D are block diagrams illustrating a sequence of steps in amethod for remote assessment of a sample assay for detection of thepresence or absence of an analyte, for example, for diagnosis of acondition of a disease, according to some embodiments. In step 300 a,enclosure 220 is provided to the user. In step 300 b, the user placesimage-capturing device 100 on a top side of enclosure 220, ensuring thataperture 225 in enclosure 220 overlaps with a lens system (e.g., opticalcouplings 115, cf. FIGS. 1A-1B) in image-capturing device 100.Alternatively, not shown in FIG. 3B, enclosure 220 comprises animage-capturing device in aperture 225 with optical couplings. In step300 c, sample cartridge 101 comprising, for example, and immunoassaytest strip, having a sensitive area 302 is introduced into the enclosure220. In an embodiment, cartridge 101 has a length and less than the fulllength of the cartridge is introduced into the enclosure. Concurrentwith or after insertion of the cartridge, according to some embodiments,light source 251 is turned on, thereby illuminating the interior ofenclosure 220. With the illuminated sample cartridge 101, the opticalcoupling of image-capturing device 100 may perform an autofocus routineso that the sensitive portion of sample cartridge 101 is well focused ona sensor array of image-capturing device 100 (e.g., sensor array 140,cf. FIG. 1A). In some embodiments, image-capturing device 100 maytolerate a slightly unfocussed image of the sample assay and samplecartridge 101, and may be configured to compensate for any blur ordistortion via image processing in an image-capturing application (e.g.,image-capturing application 122, cf. FIG. 1A). The autofocus routine isbeneficial because image-capturing device 100 may not be setupappropriately at the time when the user places it on enclosure 220 (step300 b) or, for embodiments where image-capturing device 100 is part ofenclosure 220, the autofocus routine is beneficial because the positionof the sensitive area of the cartridge may vary from one cartridge toanother. Accordingly, step 300 d includes capturing light scattered fromthe sensitive area of the sample cartridge (e.g., fluorescence light260, cf. FIG. 2) into image-capturing device 100, forming an image, andeither processing the image in image-capturing device 100, or sendingthe image to a remote server for processing and to a remote database forstorage (e.g., server 130 and database 152, cf. FIGS. 1A-1B). In someembodiments, both processing and storage functions may be performed atleast partially by the image-capturing device.

FIG. 4A illustrates an enclosure including a sample cartridge 401 havinga colored fiducial 407 a that includes a sensitive area 402, foridentifying the location of a sample assay feature, according to someembodiments. In some embodiments, an excitation-based sample assaytriggers fluorescence emission at a specified wavelength (e.g., thewavelength of the fluorescence light source 451-1, such as 385 nm, andthe like). In some embodiments, before preforming the fluorescencestimulation and analysis with the fluorescence light source 451-1, asetup step is performed to identify the location of interest, using asecond light source 451-2. Light source 451-2 may include a white lightsource or any other light source (e.g., a white LED) that includes abroadband wavelength emission range separate from the fluorescence pumpwavelength.

The setup step includes an insert-within dark chamber (or enclosure) 405to receive sample cartridge 401. Insert-within enclosure 405 has anaperture that allows optical access to the sample assay inside fromoptics coupling mechanism 415 in image-capturing device 400. Coloredfiducial 407A is printed around the assay aperture (or disposed aroundthe aperture with a sticker, and the like). Colored fiducial 407A mayhave a box shape surrounding the area of interest (the sample assay) ina specific color (such as green).

A setup step includes turning light source 451-2 ‘on,’ to illuminate theentire system. Image-capturing device 400 collects a picture ofinsert-within enclosure 405 with colored fiducial mark 407 a indicatingthe assay location. Image-capturing application 422 scans the picture tofind the location of colored fiducial 407 a. The coordinates of coloredfiducial 407 a are used to precisely locate the assay sample within aframe of the camera in optics coupling mechanism 415. In someembodiments, image-capturing application 422 may be configured to findthe corners of the green box in the colored fiducial 407 a.

When the sample assay is precisely located, light source 451-2 is turned‘off’ and fluorescent light source 451-1 is turned ‘on’ to interact withthe assay. The coordinates created with colored fiducial 407 a are usedto locate the assay and its subcomponents such as sample/control lines.

FIG. 4B illustrates insert-within box or enclosure 405 including samplecartridge 401 having a fluorescent fiducial 407 b that includes asensitive area 402, for identifying the location of a sample assayfeature, according to some embodiments. Accordingly, embodimentsconsistent with this disclosure may not use the white light source toilluminate a colored fiducial. Instead, fluorescent fiducial 407 b isformed of a fluorescent material (e.g., ink/dye and the like) that maybe excited with the same fluorescent light source that excites thesample assay. Optics coupling mechanism 415, fluorescence light source451-1, and image-capturing application 422 are as described above inreference to FIG. 4A.

In some embodiments, the emission wavelength of fluorescent fiducial 407b, while detectable by image-capturing device 400, may be different fromthe emission wavelength of the sample assay. For example, in someembodiments (e.g., when the sample assay emits red fluorescence at about650 nm), fluorescent fiducial 407 b may be selected in the greenwavelength. In such configuration, the full spectral efficiency of ared-green-blue pixel array in image-capturing device 400 (as iscustomary in smart phones and other mobile appliances) may beefficiently used.

FIG. 5 illustrates a series of images 500 a, 500 b, and 500 d(hereinafter, collectively referred to as “images 500”) of the sensitivearea of a sample cartridge including a blank assay (e.g., no teststrips), collected over time, according to some embodiments. Across-section of the blue, green, and red pixel counts 551 in thelongitudinal direction of the sample assay is also shown, for each oneof images 500. In the example illustrated, a blank assay is used (notest strip), and therefore little to no red fluorescence is detected.Most of the signal observed comes from blue scattered light form afluorescence excitation light (e.g., fluorescent excitation light 260),and as the signal progresses there are no features that arise, otherthan a ‘hump’ or step in the pixel count that indicates the startingedge of the sensitive area and the diffusion boundary of the assay. Inthe figure, the diffusion boundary 510 a, 510 b, and 510 c (hereinafter,collectively referred to as “diffusion boundary 510”) moves from left toright as illustrated by a rise in the signal for the red, blue, andgreen pixel counts, the rise indicated at dashed line 560.

In the figure, multiple images 500 of a fluorescence assay 501 areobtained at different times, t₁, t₂, and t₃, respectively. A fiducial507 indicates the position and limits of the sensitive area includingthe pixels having relevant assay information. While images 500 may notbe consecutive, in some embodiments, the sampling time, e.g., the timeinterval between times t₁, t₂, and t₃, may be 15 seconds. For each ofimages 500, the collection time, per image capture, may be as low asabout ½ second, or even less. In some embodiments, images 500 may becaptured from a video sequence (e.g., when the image-capturing deviceincludes video recording capabilities).

FIG. 6 illustrates an image 600 of a sensitive area 602 of a samplecartridge 601 including a sample assay and positioning marks 603measured with an image-capturing device (e.g., image-capturing device100, cf. FIGS. 1A-1B), according to some embodiments. In someembodiments, sensitive area 602 may include multiple test strips or testlines 611 disposed in parallel to one another, at fixed intervals, andin a perpendicular direction to a general diffusion travel 650 of theassay solution. Test lines in sensitive area 602 typically correspond toregions on an immunoassay test strip where an immobilized binding memberis positioned for capture of an indicator of the presence or absence ofan analyte of interest. Test lines in sensitive area 602 may alsoinclude control and reference lines, as is known in the area ofimmunoassay test devices. Some embodiments may include adjusting image600 by finding a scale factor between image 600 and sample cartridge601. Accordingly, positioning marks 603 indicate the edges of teststrips in the sample assay, and a proper scale factor may be determinedby comparing a number of pixels between positioning marks 603 with theknown width and positioning of test strips 611. For example, in someembodiments, positioning marks 603 may be determined by forming aspatial derivative of a pixel count function in the horizontal and/orvertical direction. The spatial derivative may include the first orsecond spatial derivative of the pixel count (∂x, ∂y, ∂²x, ∂²y, ∂x∂y).

In some embodiments, a digital filtering technique may be applied toimage 600 to remove spectral and spatial artifacts from the sensitivearea of sample cartridge 601. The spectral artifacts may include, forexample, residual fluorescence excitation light (for example on the blueside of the spectrum) that may account for some of the blue pixelvalues, or even some of the red pixel values, in the sensor array. Thismay be the case, for example, when the red pixels of the image-capturingdevice show some sensitivity to at least a portion of the blue emissionspectrum of the light emitting source.

Some embodiments extract a value for assessing a diagnostic of thesample assay by spatially filtering image 600 and spectrally filteringimage 600 as described above. Accordingly, the filtered pixel values maybe aggregated and compared to a pre-selected threshold. Thus, when theaggregated value is lower or greater than the threshold, the diseasediagnosis may be positive. Some embodiments may include error valuesbased on statistical analysis and calibration, to provide a confidenceinterval for the diagnostics.

FIG. 7 illustrates an image 700 of a sensitive area 702 of a samplecartridge 701 collected and processed by an image-capturing device(e.g., image-capturing devices 100, cf. FIGS. 1A-1B), according to someembodiments. Accordingly, a light emitting device is configured toexcite a fluorescence light from sample cartridge 701 (e.g., lightemitting source 251 and fluorescent light 260, cf. FIG. 2). In someembodiments, the fluorescence light has a wavelength within the selectedcolor in a sensor array in the image-capturing device (e.g., sensorarray 140, cf. FIG. 1A).

An immunoassay test strip contained in a sample cartridge, in someembodiments, emits fluorescence light primarily from fluorophoresattached to the target analyte, as they are fixed on the substrate byadherence to the immuno-proteins in the immunoassay strip (e.g.,adsorption, chemi-sorption, immune-ligand, and the like). Accordingly,the presence of a red light within the boundaries of sensitive area 702(e.g., an immunoassay strip) is mostly attributable to the presence ofthe target analyte (e.g., pathogens in a disease control, and the like).However, the amount of red signal within the boundaries of sensitivearea 702 may include some fluorescence background 720 a and even someexcitation light background 720 b. To better assess the backgroundsignal (e.g., not originated by target analytes fixed on the substrateof the immunoassay strip), some sample cartridges 701 may include acontrol test line 711 b, or a reference test line, in addition to testline 711 a.

FIG. 8 illustrates cross-sections 850 b, 850 r, and 850 g (hereinafter,collectively referred to as “cross-sections 850”) of fluorescencesignals captured from a sample assay along a longitudinal direction,according to some embodiments. A sample cartridge with an immunoassaycontained therein 801 includes a sensitive area 802 with test line811-1, test line 811-2, and test line 811-3 (hereinafter, collectivelyreferred to as “test lines 811”). An image-capturing application (e.g.,image-capturing application 122, cf. FIGS. 1A-1B) integrates pixelcounts along the width direction of the sample assay (along verticaldotted lines in the figure) in cross-sections 850. In some embodiments,the image-capturing application may separately integrate the values ofblue pixels (850 b), red pixels (850 r), and green pixels (850 g).Accordingly, in embodiments where the fluorescent light source is in theblue or UV spectral region (e.g., 385 nm), blue pixels (cross-section850 b) will mostly capture a background scattering from the fluorescentlight source. When this is the case, the fluorescence emission may havea wavelength in the red visible spectral region (e.g., 650 nm), and theintegration of red pixels in cross-section 850 r by the image-capturingapplication will distinctly collect the fluorescent emission from eachof the lines 811. Note how, according to some embodiments, pixelintegration may accurately capture an intensity of the fluorescentsignal emitted from each of test lines 811, thus enabling accuratequantification of the measurements. The green pixels (cross-section 850g), while not sensitive to either the fluorescent light source or thefluorescence emission, may be sensitive to a colored fiducial (e.g.,colored fiducial 407 a or fluorescent fiducial 407 b, cf. FIGS. 4A and4B), or may provide a flat background signal indicative of the profileof the sample assay (e.g., width), and the location and width of each ofthe test lines.

As shown in the figure, the three different spectral data analysis(e.g., blue pixels (850 b), red pixels (850 r), and green pixels (850g)) are collected and performed simultaneously, or almostsimultaneously, during a single image-capturing sequence. Accordingly,in some embodiments, a user may not be asked to perform any extra step,nor even to align sample cartridge 801 relative to the image-capturingdevice, as this alignment may be performed in software by theimage-capturing application using the fiducial mark.

FIG. 9 is a flow chart illustrating steps in a method 900 for capturingan image of a sensitive area in a sample cartridge using animage-capturing device and an enclosure (e.g., sensitive area 202,sample cartridge 101, image-capturing devices 100, and enclosures 120and 220, cf. FIGS. 1A-1B, and 2), according to some embodiments. Method900 may be performed at least partially by a computer in a server oruser device (e.g., server 130, user device 110, cf., FIGS. 1A-1B, 2, 3,and 4A-4B). Accordingly, at least some of the steps in method 900 may beperformed by a processor executing instructions stored in a memory andproviding data to a remote database through a network (e.g.,image-capturing application 122, database 152, and network 150, cf.FIGS. 1A and 1B). Further, methods consistent with the presentdisclosure may include at least one step as described in method 900. Insome embodiments, methods consistent with the present disclosure includeone or more steps in method 900 performed in a different order,simultaneously, almost simultaneously, or overlapping in time.

Step 902 includes inserting a sample cartridge in a dark chamber (orenclosure). In some embodiments, step 902 includes interacting a usersample containing a biological residual from a user with a reagent, inthe sample cartridge.

Step 904 includes activating a light emitter in the dark chamber. Insome embodiments, step 904 includes activating a fluorescence excitationsource.

Step 906 includes placing an image-capturing device on an aperture ofthe dark chamber.

Step 908 includes focusing an optical coupling mechanism in theimage-capturing device to form an image of a sensitive area in thesample cartridge.

Step 910 includes capturing the image of the sensitive area after aselected period of time. In some embodiments, step 910 includescapturing a second image of the sensitive area after a second period oftime, and determining the selected period of time and the second periodof time based on a diffusion time for a reagent fluid in a membranecontained in the sensitive area in the sample cartridge. In someembodiments, step 910 includes storing the image of the sensitive areain a memory in the image-capturing device. In some embodiments, step 910transmits the image of the sensitive area over a network to a remoteserver. In some embodiments, step 910 includes capturing a digital imageincluding discrete values associated with an array of pixels in theimage-capturing device, the method further comprising applying a spatialfilter and a color filter to the digital image to determine a presenceof a target agent in the sensitive area of the sample cartridge. In someembodiments, step 910 includes determining the selected period of timeaccording to a threshold of an intensity of a red light captured by theimage-capturing device from a selected portion of the sensitive area. Insome embodiments, step 910 includes storing the image as a fluorescencebackground image for data processing.

FIG. 10 is a flow chart illustrating steps in a computer-implementedmethod 1000 for remotely diagnosing a disease or condition, or fordetecting presence or absence of an analyte or biomarker, from an imageof the sensitive area in a sample cartridge captured with animage-capturing device (e.g., sensitive area 202, sample cartridge 101,image-capturing devices 100, and enclosures 120 and 220, cf. FIGS.1A-1B, and 2), according to some embodiments. The sample cartridgecomprises or corresponds to an immunoassay test. Method 1000 may beperformed at least partially by a computer in a server or client (user)device (e.g., server 130, client device 110, cf., FIGS. 1A-1B, 2, 3, and4A-4B). Accordingly, at least some of the steps in method 1000 may beperformed by a processor executing instructions stored in a memory andproviding data to a remote database through a network (e.g.,image-capturing application 122, database 152, and network 150, cf.FIGS. 1A and 1B). Further, methods consistent with the presentdisclosure may include at least one step as described in method 1000. Insome embodiments, methods consistent with the present disclosure includeone or more steps in method 1000 performed in a different order,simultaneously, almost simultaneously, or overlapping in time.

Step 1002 includes identifying, upon receipt of a user input, afiduciary figure in a bottom side of an enclosure with animage-capturing device.

Step 1004 includes adjusting an optical coupling in the image-capturingdevice to obtain a sharp image of the fiduciary figure.

Step 1006 includes identifying a sensitive area of a sample cartridgewithin a field of view of the optical coupling.

Step 1008 includes adjusting an image of the sensitive area of thesample cartridge. In some embodiments, step 1008 includes removing ahorizontal skew and a vertical skew in the image. In some embodiments,step 1008 includes finding a border of the sensitive area of the samplecartridge and applying geometrical transformations on an area delimitedby the border of the sample cartridge. In some embodiments, step 1008includes filtering the image of the sensitive area of the samplecartridge to remove a color and a spatial artifact from the sensitivearea of the sample cartridge. In some embodiments, step 1008 includesfinding a scale factor between the image of the sensitive area of thesample cartridge and the sample cartridge.

Step 1010 includes identifying a target region within the image of thesensitive area of the sample cartridge.

Step 1012 includes extracting a value of a selected color for multiplepixels in the target region. In some embodiments, step 1012 includesspatially filtering the image and spectrally filtering the image toobtain a selected group of pixel values to form the value of theselected color.

Step 1014 includes determining a presence of a target analyte when thevalue of the selected color is greater than a pre-selected threshold.

FIG. 11 is a flow chart illustrating steps in a method 1100 for remotelydiagnosing a disease or condition, or for detecting presence or absenceof an analyte or biomarker, with an image-capturing device, according tosome embodiments. Method 1100 may be performed at least partially by acomputer in a server or client device (e.g., server 130, client device110, cf., FIGS. 1A-1B, 2, 3, and 4A-4B). Accordingly, at least some ofthe steps in method 1100 may be performed by a processor executinginstructions stored in a memory and providing data to a remote databasethrough a network (e.g., image-capturing application 122, database 152,and network 150, cf. FIGS. 1A and 1B). Further, methods consistent withthe present disclosure may include at least one step as described inmethod 1100. In some embodiments, methods consistent with the presentdisclosure include one or more steps in method 1100 performed in adifferent order, simultaneously, almost simultaneously, or overlappingin time.

Step 1102 includes inserting a sample cartridge in a dark chamber. Insome embodiments, step 1102 includes interacting a user samplecontaining a biological residual from a user with a reagent, in thesample cartridge. In some embodiments, step 1102 includes placing thesample cartridge in an insert-within box in the dark chamber.

Step 1104 includes activating a fluorescent excitation light (e.g., ablue LED, or a UV LED emitting at 385 nm) in the dark chamber. In someembodiments, step 1104 includes activating a fluorescence excitationsource.

Step 1106 includes placing an image-capturing device on an aperture ofthe dark chamber, the image-capturing device including filters toimprove measurement specificity and to reduce background.

Step 1108 includes using a colored fluorescent fiducial to determine anarea of interest in the sample cartridge. In some embodiments, the greenfluorescent fiducial is marked, or printed, or attached to a surface inthe insert-within box of the dark chamber.

Step 1110 includes running a sample assay. In some embodiments, step1110 includes causing the sample assay to diffuse from one edge of asensitive area in the sample cartridge to the other edge of thesensitive area along a longitudinal direction of the sample cartridge.

Step 1112 includes collecting images periodically, with theimage-capturing device, as the sample assay progresses (e.g., viacapillary diffusion) to the end of the sensitive area in the samplecartridge. In some embodiments, step 1112 includes collecting imagesevery 10 to 20 seconds, or every 15 seconds. In some embodiments, step1112 may include collecting a continuous video of the sample assay as itprogresses to the end of the sensitive area in the sample cartridge. Insome embodiments, step 1112 includes transmitting the collected image orvideo to a remote server for processing. Further, in some embodiments,step 1112 includes processing the collected image or video with animage-capturing application in the image-capturing device, and storingthe collected image in a memory in the image-capturing device. Further,in some embodiments, step 1112 includes analyzing all or a portion of animage or video to obtain a result, and transmitting the result over anetwork, for example, to a remote server or a database.

FIG. 12 is a flow chart illustrating steps in a method 1200 fordiagnosing a disease or condition, or for detecting presence or absenceof an analyte or biomarker, with an image-capturing device, according toother embodiments. Method 1200 may be performed at least partially by acomputer in a server or client (user) device (e.g., server 130, clientdevice 110, cf., FIGS. 1A-1B, 2, 3, and 4A-4B). Accordingly, at leastsome of the steps in method 1200 may be performed by a processorexecuting instructions stored in a memory and providing data to a remotedatabase through a network (e.g., image-capturing application 122,database 152, and network 150, cf. FIGS. 1A and 1B). Further, methodsconsistent with the present disclosure may include at least one step asdescribed in method 1200. In some embodiments, methods consistent withthe present disclosure include one or more steps in method 1200performed in a different order, simultaneously, almost simultaneously,or overlapping in time.

Step 1202 includes inserting a sample cartridge in a dark chamber orenclosure.

Step 1204 includes activating a light emitter in the dark chamber orenclosure.

Step 1206 includes focusing an optical coupling mechanism in animage-capturing device to optimize an image of a sensitive area in thesample cartridge.

Step 1208 includes capturing, with an image-capturing device, an imageof a sensitive area in the sample cartridge after a selected period oftime. In some embodiments, step 1208 includes at least one of: (i)capturing multiple images of the sensitive area at a selectedperiodicity, or (ii) capturing a video recording of a sample assay as itis diffusively transported along the sensitive area in the samplecartridge. The step may optionally include identifying the sensitivearea in the sample cartridge based on a shape and location of thefluorescent fiducial mark. It will be appreciated that subsequent tocapturing one or more images or a video the one or more images or avideo may be analyzed, before or after transmitting to a remote serveror database. In some embodiments, Step 1208 includes analyzing one ormore images or a video before transmitting to the remote server ordatabase.

Step 1210 includes providing the image of the sensitive area to aprocessor, wherein the processor comprises an image-capturingapplication. In one embodiment, an analysis of an image before it isprovided to the image-capturing application my occur. Such an analysiscan be an analysis of a portion of an image or a partial of an image.

FIG. 13 is a flow chart illustrating steps in a method 1300 fordiagnosing a disease or condition, or for detecting presence or absenceof an analyte or biomarker, with an image-capturing device, according toother embodiments. Method 1300 may be performed at least partially by acomputer in a server or client device (e.g., server 130, client device110, cf., FIGS. 1A-1B, 2, 3, and 4A-4B). Accordingly, at least some ofthe steps in method 1300 may be performed by a processor executinginstructions stored in a memory and providing data to a remote databasethrough a network (e.g., image-capturing application 122, database 152,and network 150, cf. FIGS. 1A and 1B). Further, methods consistent withthe present disclosure may include at least one step as described inmethod 1300. In some embodiments, methods consistent with the presentdisclosure include one or more steps in method 1300 performed in adifferent order, simultaneously, almost simultaneously, or overlappingin time.

Step 1302 includes identifying, upon receipt of a user input, afiduciary figure in a bottom side of an enclosure with animage-capturing device.

Step 1304 includes adjusting an optical coupling in the image-capturingdevice to obtain a sharp image of the fiduciary figure. In someembodiments, step 1304 further includes removing a horizontal skew and avertical skew to adjust an image of the sensitive area of the samplecartridge. In some embodiments, step 1304 includes filtering an image ofthe sensitive area of the sample cartridge to remove a color and aspatial artifact from the sensitive area of the sample cartridge. Insome embodiments, step 1304 includes finding a scale factor between theimage of the sensitive area of the sample cartridge and the samplecartridge.

Step 1306 includes identifying a sensitive area of a sample cartridgewithin a field of view of the optical coupling.

Step 1308 includes finding a border of the sensitive area of the samplecartridge and applying geometrical transformations on an area delimitedby the border of the sample cartridge.

Step 1310 includes identifying a target region within the sensitive areaof the sample cartridge.

Step 1312 includes extracting a value of a selected color for multiplepixels in the target region.

Step 1314 includes determining a presence of a target analyte when thevalue of the selected color is greater than a pre-selected threshold.

FIG. 14 is a block diagram illustrating an example computer system 1400with which the image-capturing device and the server of FIG. 1, and themethods as disclosed herein can be implemented, according to someembodiments. In certain aspects, computer system 1400 may be implementedusing hardware or a combination of software and hardware, either in adedicated server, or integrated into another entity, or distributedacross multiple entities.

Computer system 1400 (e.g., server in FIG. 1) includes a bus 1408 orother communication mechanism for communicating information, and aprocessor 1402 coupled with bus 1408 for processing information. By wayof example, computer system 1400 may be implemented with one or moreprocessors 1402. Processor 1402 may be a general-purpose microprocessor,a microcontroller, a Digital Signal Processor (DSP), an ApplicationSpecific Integrated Circuit (ASIC), a Field Programmable Gate Array(FPGA), a Programmable Logic Device (PLD), a controller, a statemachine, gated logic, discrete hardware components, or any othersuitable entity that can perform calculations or other manipulations ofinformation.

Computer system 1400 can include, in addition to hardware, code thatcreates an execution environment for the computer program in question,e.g., code that constitutes processor firmware, a protocol stack, adatabase management system, an operating system, or a combination of oneor more of them stored in an included memory, such as a Random AccessMemory (RAM), a flash memory, a Read-Only Memory (ROM), a ProgrammableRead-Only Memory (PROM), an Erasable PROM (EPROM), registers, a harddisk, a removable disk, a CD-ROM, a DVD, or any other suitable storagedevice, coupled to the bus for storing information and instructions tobe executed by the processor. Processor 1402 and memory 1404 can besupplemented by, or incorporated in, special purpose logic circuitry.

The instructions may be stored in memory 1404 and implemented in one ormore computer program products, i.e., one or more modules of computerprogram instructions encoded on a computer-readable medium for executionby, or to control the operation of, the computer system, and accordingto any method well-known to those of skill in the art, including, butnot limited to, computer languages such as data-oriented languages(e.g., SQL, dBase), system languages (e.g., C, Objective-C, C++,Assembly), architectural languages (e.g., Java, .NET), and applicationlanguages (e.g., PHP, Ruby, Perl, Python). Instructions may also beimplemented in computer languages such as array languages,aspect-oriented languages, assembly languages, authoring languages,command-line interface languages, compiled languages, concurrentlanguages, curly-bracket languages, dataflow languages, data-structuredlanguages, declarative languages, esoteric languages, extensionlanguages, fourth-generation languages, functional languages,interactive-mode languages, interpreted languages, iterative languages,list-based languages, little languages, logic-based languages, machinelanguages, macro languages, metaprogramming languages, multiparadigmlanguages, numerical analysis, non-English-based languages,object-oriented class-based languages, object-oriented prototype-basedlanguages, off-side rule languages, procedural languages, reflectivelanguages, rule-based languages, scripting languages, stack-basedlanguages, synchronous languages, syntax handling languages, visuallanguages, wirth languages, and xml-based languages. The memory may alsobe used for storing temporary variable or other intermediate informationduring execution of instructions to be executed by the processor.

A computer program as discussed herein does not necessarily correspondto a file in a file system. A program can be stored in a portion of afile that holds other programs or data (e.g., one or more scripts storedin a markup language document), in a single file dedicated to theprogram in question, or in multiple coordinated files (e.g., files thatstore one or more modules, subprograms, or portions of code). A computerprogram can be deployed to be executed on one computer or on multiplecomputers that are located at one site or distributed across multiplesites and interconnected by a communication network. The processes andlogic flows described in this specification can be performed by one ormore programmable processors executing one or more computer programs toperform functions by operating on input data and generating output.

Computer system 1400 further includes a data storage device 1406 such asa magnetic disk or optical disk, coupled to the bus for storinginformation and instructions. Computer system 1400 may be coupled via aninput/output module 1410 to various devices. Input/output module 1410can be any input/output module. Exemplary input/output modules includedata ports such as USB ports. Input/output module 1410 is configured toconnect to a communications module 1412. Exemplary communicationsmodules include networking interface cards, such as Ethernet cards andmodems. In certain aspects, input/output module 1410 may be configuredto connect to a plurality of devices, such as an input device 1414and/or an output device 1416. Exemplary input devices include a keyboardand a pointing device, e.g., a mouse or a trackball, by which a user canprovide input to the computer system. Other kinds of input devices canbe used to provide for interaction with a user as well, such as atactile input device, visual input device, audio input device, orbrain-computer interface device. For example, feedback provided to theuser can be any form of sensory feedback, e.g., visual feedback,auditory feedback, or tactile feedback; and input from the user can bereceived in any form, including acoustic, speech, tactile, or brain waveinput. Exemplary output devices include display devices, such as an LCD(liquid crystal display) monitor for displaying information to the user.

In some embodiments, computer system 1400 is a network-based,voice-activated device accessed by the user. Input/output device 1414 or1416 may include a microphone providing the queries in voice format, andreceiving multiple inputs from the user, also in a voice format, in thelanguage of the user. Further, in some embodiments, a neural linguisticalgorithm may cause the voice-activated device to contact the user backand receive a user selection of the respiratory mask via a voice commandor request.

According to one aspect of the present disclosure, the image-capturingdevice and server of FIG. 1A can be implemented using computer system1400 in response to processor 1402 executing one or more sequences ofone or more instructions contained in memory 1404. Such instructions maybe read into the memory from another machine-readable medium, such asthe data storage device. Execution of the sequences of instructionscontained in the main memory causes processor 1402 to perform theprocess steps described herein. One or more processors in amulti-processing arrangement may also be employed to execute thesequences of instructions contained in the memory. In alternativeaspects, hard-wired circuitry may be used in place of or in combinationwith software instructions to implement various aspects of the presentdisclosure. Thus, aspects of the present disclosure are not limited toany specific combination of hardware circuitry and software.

Various aspects of the subject matter described in this specificationcan be implemented in a computing system that includes a back-endcomponent, e.g., as a data server, or that includes a middlewarecomponent, e.g., an application server, or that includes a front-endcomponent, e.g., an image-capturing device having a graphical userinterface or a Web browser through which a user can interact with animplementation of the subject matter described in this specification, orany combination of one or more such back-end, middleware, or front-endcomponents. The components of the system can be interconnected by anyform or medium of digital data communication, e.g., a communicationnetwork. The communication network (e.g., the network in FIG. 1) caninclude, for example, any one or more of a LAN, a WAN, the Internet, andthe like. Further, the communication network can include, but is notlimited to, for example, any one or more of the following networktopologies, including a bus network, a star network, a ring network, amesh network, a star-bus network, tree or hierarchical network, or thelike. The communications modules can be, for example, modems or Ethernetcards.

Computer system 1400 can include image-capturing devices and serverswherein the image-capturing device and server are generally remote fromeach other and typically interact through a communication network. Therelationship of image-capturing device and server arises by virtue ofcomputer programs running on the respective computers and having animage-capturing device-server relationship to each other. Computersystem 1400 can be, for example, and without limitation, a desktopcomputer, laptop computer, or tablet computer. Computer system 1400 canalso be embedded in another device, for example, and without limitation,a mobile telephone, a PDA, a mobile audio player, a Global PositioningSystem (GPS) receiver, a video game console, and/or a television set topbox.

The term “machine-readable storage medium” or “computer-readable medium”as used herein refers to any medium or media that participates inproviding instructions to the processor for execution. Such a medium maytake many forms, including, but not limited to, non-volatile media,volatile media, and transmission media. Non-volatile media include, forexample, optical or magnetic disks, such as the data storage device.Volatile media include dynamic memory, such as the memory. Transmissionmedia include coaxial cables, copper wire, and fiber optics, includingthe wires that comprise the bus. Common forms of machine-readable mediainclude, for example, floppy disk, a flexible disk, hard disk, magnetictape, any other magnetic medium, a CD-ROM, DVD, any other opticalmedium, punch cards, paper tape, any other physical medium with patternsof holes, a RAM, a PROM, an EPROM, a FLASH EPROM, any other memory chipor cartridge, or any other medium from which a computer can read. Themachine-readable storage medium can be a machine-readable storagedevice, a machine-readable storage substrate, a memory device, acomposition of matter effecting a machine-readable propagated signal, ora combination of one or more of them.

As used herein, the phrase “at least one of” preceding a series ofitems, with the terms “and” or “or” to separate any of the items,modifies the list as a whole, rather than each member of the list (e.g.,each item). The phrase “at least one of” does not require selection ofat least one item; rather, the phrase allows a meaning that includes atleast one of any one of the items, and/or at least one of anycombination of the items, and/or at least one of each of the items. Byway of example, the phrases “at least one of A, B, and C” or “at leastone of A, B, or C” each refer to only A, only B, or only C; anycombination of A, B, and C; and/or at least one of each of A, B, and C.

To the extent that the term “include,” “have,” or the like is used inthe description or the claims, such term is intended to be inclusive ina manner similar to the term “comprise” as “comprise” is interpretedwhen employed as a transitional word in a claim. The word “exemplary” isused herein to mean “serving as an example, instance, or illustration.”Any embodiment described herein as “exemplary” is not necessarily to beconstrued as preferred or advantageous over other embodiments.

A reference to an element in the singular is not intended to mean “oneand only one” unless specifically stated, but rather “one or more.” Allstructural and functional equivalents to the elements of the variousconfigurations described throughout this disclosure that are known orlater come to be known to those of ordinary skill in the art areexpressly incorporated herein by reference and intended to beencompassed by the subject technology. Moreover, nothing disclosedherein is intended to be dedicated to the public regardless of whethersuch disclosure is explicitly recited in the above description.

While this specification contains many specifics, these should not beconstrued as limitations on the scope of what may be claimed, but ratheras descriptions of particular implementations of the subject matter.Certain features that are described in this specification in the contextof separate embodiments can also be implemented in combination in asingle embodiment. Conversely, various features that are described inthe context of a single embodiment can also be implemented in multipleembodiments separately or in any suitable subcombination. Moreover,although features may be described above as acting in certaincombinations and even initially claimed as such, one or more featuresfrom a claimed combination can in some cases be excised from thecombination, and the claimed combination may be directed to asubcombination or variation of a subcombination.

The subject matter of this specification has been described in terms ofparticular aspects, but other aspects can be implemented and are withinthe scope of the following claims. For example, while operations aredepicted in the drawings in a particular order, this should not beunderstood as requiring that such operations be performed in theparticular order shown or in sequential order, or that all illustratedoperations be performed, to achieve desirable results. The actionsrecited in the claims can be performed in a different order and stillachieve desirable results. As one example, the processes depicted in theaccompanying figures do not necessarily require the particular ordershown, or sequential order, to achieve desirable results. In certaincircumstances, multitasking and parallel processing may be advantageous.Moreover, the separation of various system components in the aspectsdescribed above should not be understood as requiring such separation inall aspects, and it should be understood that the described programcomponents and systems can generally be integrated together in a singlesoftware product or packaged into multiple software products. Othervariations are within the scope of the following claims.

In one aspect, a method may be an operation, an instruction, or afunction and vice versa. In one aspect, a claim may be amended toinclude some or all of the words (e.g., instructions, operations,functions, or components) recited in other one or more claims, one ormore words, one or more sentences, one or more phrases, one or moreparagraphs, and/or one or more claims.

To illustrate the interchangeability of hardware and software, itemssuch as the various illustrative blocks, modules, components, methods,operations, instructions, and algorithms have been described generallyin terms of their functionality. Whether such functionality isimplemented as hardware, software, or a combination of hardware andsoftware depends upon the particular application and design constraintsimposed on the overall system. Skilled artisans may implement thedescribed functionality in varying ways for each particular application.

As used herein, the phrase “at least one of” preceding a series ofitems, with the terms “and” or “or” to separate any of the items,modifies the list as a whole, rather than each member of the list (e.g.,each item). The phrase “at least one of” does not require selection ofat least one item; rather, the phrase allows a meaning that includes atleast one of any one of the items, and/or at least one of anycombination of the items, and/or at least one of each of the items. Byway of example, the phrases “at least one of A, B, and C” or “at leastone of A, B, or C” each refer to only A, only B, or only C; anycombination of A, B, and C; and/or at least one of each of A, B, and C.

The word “exemplary” is used herein to mean “serving as an example,instance, or illustration.” Any embodiment described herein as“exemplary” is not necessarily to be construed as preferred oradvantageous over other embodiments. Phrases such as an aspect, theaspect, another aspect, some aspects, one or more aspects, animplementation, the implementation, another implementation, someimplementations, one or more implementations, an embodiment, theembodiment, another embodiment, some embodiments, one or moreembodiments, a configuration, the configuration, another configuration,some configurations, one or more configurations, the subject technology,the disclosure, the present disclosure, other variations thereof andalike are for convenience and do not imply that a disclosure relating tosuch phrase(s) is essential to the subject technology or that suchdisclosure applies to all configurations of the subject technology. Adisclosure relating to such phrase(s) may apply to all configurations,or one or more configurations. A disclosure relating to such phrase(s)may provide one or more examples. A phrase such as an aspect or someaspects may refer to one or more aspects and vice versa, and thisapplies similarly to other foregoing phrases.

A reference to an element in the singular is not intended to mean “oneand only one” unless specifically stated, but rather “one or more.”Pronouns in the masculine (e.g., his) include the feminine and neutergender (e.g., her and its) and vice versa. The term “some” refers to oneor more. Underlined and/or italicized headings and subheadings are usedfor convenience only, do not limit the subject technology, and are notreferred to in connection with the interpretation of the description ofthe subject technology. Relational terms such as first and second andthe like may be used to distinguish one entity or action from anotherwithout necessarily requiring or implying any actual such relationshipor order between such entities or actions. All structural and functionalequivalents to the elements of the various configurations describedthroughout this disclosure that are known or later come to be known tothose of ordinary skill in the art are expressly incorporated herein byreference and intended to be encompassed by the subject technology.Moreover, nothing disclosed herein is intended to be dedicated to thepublic regardless of whether such disclosure is explicitly recited inthe above description. No claim element is to be construed under theprovisions of 35 U.S.C. § 112, sixth paragraph, unless the element isexpressly recited using the phrase “means for” or, in the case of amethod claim, the element is recited using the phrase “step for.”

While this specification contains many specifics, these should not beconstrued as limitations on the scope of what may be described, butrather as descriptions of particular implementations of the subjectmatter. Certain features that are described in this specification in thecontext of separate embodiments can also be implemented in combinationin a single embodiment. Conversely, various features that are describedin the context of a single embodiment can also be implemented inmultiple embodiments separately or in any suitable subcombination.Moreover, although features may be described above as acting in certaincombinations and even initially described as such, one or more featuresfrom a described combination can in some cases be excised from thecombination, and the described combination may be directed to asubcombination or variation of a subcombination.

The subject matter of this specification has been described in terms ofparticular aspects, but other aspects can be implemented and are withinthe scope of the following claims. For example, while operations aredepicted in the drawings in a particular order, this should not beunderstood as requiring that such operations be performed in theparticular order shown or in sequential order, or that all illustratedoperations be performed, to achieve desirable results. The actionsrecited in the claims can be performed in a different order and stillachieve desirable results. As one example, the processes depicted in theaccompanying figures do not necessarily require the particular ordershown, or sequential order, to achieve desirable results. In certaincircumstances, multitasking and parallel processing may be advantageous.Moreover, the separation of various system components in the aspectsdescribed above should not be understood as requiring such separation inall aspects, and it should be understood that the described programcomponents and systems can generally be integrated together in a singlesoftware product or packaged into multiple software products.

The title, background, brief description of the drawings, abstract, anddrawings are hereby incorporated into the disclosure and are provided asillustrative examples of the disclosure, not as restrictivedescriptions. It is submitted with the understanding that they will notbe used to limit the scope or meaning of the claims. In addition, in thedetailed description, it can be seen that the description providesillustrative examples and the various features are grouped together invarious implementations for the purpose of streamlining the disclosure.The method of disclosure is not to be interpreted as reflecting anintention that the described subject matter requires more features thanare expressly recited in each claim. Rather, as the claims reflect,inventive subject matter lies in less than all features of a singledisclosed configuration or operation. The claims are hereby incorporatedinto the detailed description, with each claim standing on its own as aseparately described subject matter.

The claims are not intended to be limited to the aspects describedherein, but are to be accorded the full scope consistent with thelanguage claims and to encompass all legal equivalents. Notwithstanding,none of the claims are intended to embrace subject matter that fails tosatisfy the requirements of the applicable patent law, nor should theybe interpreted in such a way.

1. A method, comprising: inserting a sample cartridge in a dark chamber;activating a light emitter in the dark chamber; positioning an opticalcoupling mechanism in an image-capturing device with respect to thesample cartridge to optimize quality of an image of a sensitive area inthe sample cartridge; capturing, with the image capturing device, animage of a sensitive area in the sample cartridge after a selectedperiod of time; providing the image of the sensitive area to aprocessor, wherein the processor comprises an image-capturingapplication; and integrating, with the image-capturing application,multiple pixel counts separately for different color pixels in a sensorarray of the image-capturing device.
 2. The method of claim 1, furthercomprising interacting a user sample containing a biological residualfrom a user with a reagent, in the sample cartridge.
 3. The method ofclaim 1, wherein activating the light emitter in the dark chambercomprises activating a fluorescence excitation source.
 4. The method ofclaim 1, further comprising capturing a second image of the sensitivearea after a second period of time, and determining the selected periodof time and the second period of time based on a diffusion time for areagent fluid in a membrane contained in the sensitive area in thesample cartridge.
 5. The method of claim 1, wherein providing the imageof the sensitive area to the processor comprises storing the image ofthe sensitive area in a memory of the image capturing device, andtransmitting the image of the sensitive area over a network to a remoteserver or to a database.
 6. The method of claim 1, wherein capturing theimage of the sensitive area comprises capturing a digital imageincluding discrete values associated with an array of pixels in theimage-capturing device, the method further comprising applying a spatialfilter and a color filter to the digital image to determine a presenceof a target agent in the sensitive area of the sample cartridge.
 7. Themethod of claim 1, further comprising determining the selected period oftime according to a threshold of an intensity of a light captured by theimage-capturing device from a control portion of the sensitive area. 8.The method of claim 1, wherein capturing the image of the sensitive areain the image-capturing device comprises storing the image as afluorescence background image.
 9. The method of claim 1, wherein usingan image-capturing device to capture an image of a sensitive area in thesample cartridge comprises capturing an image of a fluorescent fiducialmark excited by the light emitter, and identifying the sensitive area inthe sample cartridge based on a shape and location of the fluorescentfiducial mark.
 10. The method of claim 1, wherein capturing the image ofthe sensitive area after a selected period of time comprises one of: (i)capturing multiple images of the sensitive area at a selectedperiodicity, or (ii) capturing a video recording of a sample assay as itis diffusively transported along the sensitive area in the samplecartridge.
 11. A system, comprising: an enclosure including a darkchamber, the enclosure configured to block ambient light from enteringthe dark chamber; a cartridge aperture on a side of the enclosure toenable a sample cartridge to be disposed at least partially inside thedark chamber, wherein the cartridge aperture is configured to at leastpartially close when the sample cartridge is at least partially disposedinside the dark chamber; a light emitting device in an interior portionof the enclosure, the light emitting device configured to emit afluorescence excitation light directed to the sample cartridge; anoptical coupler inside the enclosure to form a partial image of thesample cartridge in a sensor array of an image-capturing device, whenthe sample cartridge is at least partially disposed in the dark chamber;and an image-capturing application configured to integrate multiplepixel counts separately for different color pixels in a sensor array ofthe image-capturing device.
 12. The system of claim 11, furthercomprising a battery configured to provide a power to the light emittingdevice, and a circuit coupling the battery with the light emittingdevice, the circuit configured to remain open when the sample cartridgeis outside the enclosure and to be closed when the sample cartridge isdisposed at least partially in the dark chamber.
 13. The system of claim11, wherein the sample cartridge comprises a focusing mark on theinterior portion, the focusing mark configured to allow theimage-capturing device to focus the optical coupler when the lightemitting device is turned ‘on’.
 14. The system of claim 11, wherein thelight emitting device in the enclosure comprises a fluorescenceexcitation source.
 15. The system of claim 11, wherein the lightemitting device is configured to excite one or more fluorescencelight(s) from the sample cartridge, and wherein the emitted fluorescencelight from the cartridge has a wavelength within a selected color in asensor array in the image-capturing device.
 16. A computer-implementedmethod, comprising: identifying, upon receipt of a user input, afiduciary figure in a bottom side of an enclosure with animage-capturing device; adjusting, in the image-capturing device, anoptical coupling to obtain a sharp image of the fiduciary figure;identifying a sensitive area of a sample cartridge within a field ofview of the optical coupling; finding a border of the sensitive area ofthe sample cartridge and applying geometrical transformations on an areadelimited by the border of the sample cartridge; identifying a targetregion within the sensitive area of the sample cartridge; integrating,with the image-capturing application, multiple pixel counts separatelyfor different color pixels in a sensor array of the image-capturingdevice; extracting a value of a selected color for multiple pixels inthe target region; and determining a presence of a target analyte whenthe value of the selected color is greater than a pre-selectedthreshold.
 17. The computer-implemented method of claim 16, furthercomprising removing a horizontal skew and a vertical skew to adjust animage of the sensitive area of the sample cartridge.
 18. Thecomputer-implemented method of claim 16, wherein adjusting an opticalcoupling comprises filtering an image of the sensitive area of thesample cartridge to remove a color and a spatial artifact from thesensitive area of the sample cartridge.
 19. The computer-implementedmethod of claim 16, wherein adjusting an optical coupling comprisesfinding a scale factor between an image of the sensitive area of thesample cartridge and the sample cartridge.
 20. The computer-implementedmethod of claim 16, wherein extracting a value comprises spatiallyfiltering an image and spectrally filtering the image to obtain aselected group of pixel values to form the value of the selected color.